We narrowed to 629 results for: Dos
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TypeBlog PostPublishedJune 18, 2020, 1:15 p.m....coronaviruses and enable the virus to enter host cells. To do this, they injected the spike protein from two different...effectively to protein targets, similar to how antibodies do. The structure of the common antibody is heftier ...
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A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog PostPublishedMay 7, 2020, 1:15 p.m....guanines). This limits the use of ABEs as DNA sites that do not contain a suitable PAM cannot be edited efficiently...While base editors are precise molecular tools, they do have undesired off-target editing effects. ABEs have... -
A Guide to Starting Your Own Journal Club
TypeBlog PostPublishedFeb. 6, 2020, 3 p.m....Choosing a topic and gauging interest The first thing to do if you’ve thought about starting a journal club is...make. How often will you hold it? How many people do you expect to attend? What is the expectation on ... -
Pooled Library Amplifications
TypeBlog PostPublishedMay 30, 2024, 1:30 p.m.... = 500,000 colonies. Pro tip! Since you probably do not want to individually count 500,000 colonies, ...did not result in significant recombination. If you do see recombination, we recommend ensuring the cells... -
Twenty Years of Addgene Sharing: CRISPR
TypeBlog PostPublishedApril 18, 2024, 1:15 p.m....use But exactly what kind of editing are people doing with all these CRISPR plasmids (and libraries)? ...scientists use CRISPR technology? We think so — and so do our requestors! CRISPR plasmids in the Addgene repository... -
AAV Vector Quality Control: Going the Extra Mile with NGS
TypeBlog PostPublishedSept. 12, 2017, 1:44 p.m....they are present at a very low concentration. So how do we make sure to catch all of these potential mishaps...expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...users titrate their AAV vectors to find the optimal dose that allows for sufficient transgene expression ... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog PostPublishedSept. 6, 2016, 2:30 p.m....proteins to pathways - Ras as a model When mutations do arise and cancer ensues, traditional chemotherapies...and thousands of investigators studying them, how do we ensure that the research community has access ... -
Which Fluorescent Protein Should I Use?
TypeBlog PostPublishedMay 20, 2014, 2:06 p.m....Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how fluorescent proteins glow? ..., you will not be able to use far red-FPs. If you do not have a filter that will pass blue light to the... -
Viral Vectors 101: The Retroviral Lifecycle
TypeBlog PostPublishedJuly 27, 2023, 1:15 p.m....cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular biologists working with ...in order to understand what these vector particles do once they come in contact with target cells (Fig.... -
Getting to Know Expansion Microscopy
TypeBlog PostPublishedJune 20, 2023, 1:15 p.m....fluorescence microscopy (Dani, et al., 2010). How do you overcome this resolution limit? Electron Microscopy...Prakash K, et al. 2022). So what’s a researcher to do if they want to understand the ultrastructural relationships... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m....gradients, practice making the iodixanol layers before doing your first purification. This helped me get a feel...to learn more about different ways to titer AAV. Do you have any AAV production tips or tricks? Please...multiplicity of infections (MOIs), to determine the optimal dose of AAV for my cells of interest. Every batch of ... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog PostPublishedOct. 4, 2018, 12:44 p.m....interference), and CRISRPa (activation) libraries that do just that (Doench et al., 2016, Sanson et al., 2018... identifying strong hits in genetic screens, they do have several limitations. CRISPRko results in a complete... -
Antibodies 101: Immunoprecipitation
TypeBlog PostPublishedDec. 7, 2021, 2:15 p.m.... beads are particularly advantageous because they do not require centrifugation between washes which can...controls include knockout cell lines or tissues that do not express the protein. With this control set, you... -
To Codon Optimize or Not: That is the Question
TypeBlog PostPublishedNov. 12, 2020, 2:15 p.m....offer tools to help with these decisions. So when do you want to codon optimize your gene of interest?...useful in expressing functional proteins in hosts that do not naturally express that gene. The protein you ... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....negative control to determine whether your PIPs actually do interact. Choosing the right negative control is ...scenarios that call for a simple, yes-no answer — i.e., do two proteins interact or not? — FRET is preferable...endogenous PIPs should decrease the BiFC signal in a dose-dependent manner. This is straightforward in easy-to-transfect...easy-to-transfect cell lines where plasmid dosage can be tightly controlled. This is also theoretically... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....but orthologs like LwaCas13a do not require a specific PFS. Cas13 enzymes do not contain the RuvC and HNH... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....time, when scientists who support open science, but do not share their own data - which seems to be the ...members or colleagues: How much unpublished AAV data do you have sitting in computers and notebooks right... gene transfer to the rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy... -
Plasmids 101: Shuttle Vectors
TypeBlog PostPublishedMay 20, 2025, 1:15 p.m....life. That’s convenient for researchers who want to do experiments in complex eukaryotic cells, but would...to express your protein of interest. However, you do still need a species-compatible promoter for expressing... -
Degrading DNA with Cascade-Cas3
TypeBlog PostPublishedFeb. 11, 2025, 2:15 p.m....five of these Cas proteins, while type I-C Cascades do not contain Cas6. Regardless of the subtype, the ...to the multi-Cas mechanism. These benefits of Cas3 do come at a cost — the previously mentioned issues ... -
Antibodies 101: The Four ELISAs and When to Use Them
TypeBlog PostPublishedJuly 25, 2024, 1:15 p.m....components can interfere with conjugates. Make sure to do your research before conjugating at the bench. ...to distinct, non-overlapping epitopes so that they do not interfere with each other. The detection antibody...